Anaerobe jar closure assembly



Dec. 9, 1969 J. H. BREWER ANAEROBE JAR CLOSURE ASSEMBLY 2 Sheets-Sheet lFiled May 31, 1966 INVENTOR. Jo/{M 1/- akin 2 Arrow/5X5 Dec. 9-, 1969 J.H. BREWER 3,483,089

ANAEROBE JAR CLOSURE ASSEMBLY Filed May 31, 1966 4 2 Sheets-Sheet 2 8.INVENTOR Jaw/v 1/. 3 511036 F G 5 BY ATTORN United States Patent3,483,089 ANAEROBE JAR CLOSURE ASSEMBLY John H. Brewer, Towson, Md.,assignor to B-D Laboratories, Inc., East Rutherford, N.J., a corporationof Delaware Filed May 31, 1966, Ser. No. 553,823 Int. Cl. C12b 1/10 US.Cl. 195127 2 Claims ABSTRACT OF THE DISCLOSURE An improved cover for ananaerobe jar assembly adapted to be mounted on the open end of theanaerobe jar in sealing engagement therewith. A clamping device whichincludes a clamping strap of greater length than the diameter of theopen end of the jar and which extends across the cover and is formedwith depending end portions with means for engaging beneath the exteriorof the flange of the jar and a clamping screw extending through thestrap in threaded engagement therewith to engage the top surface of thecover so as to releasably hold the cover in place on the jar.

This invention relates to an improved anaerobe jar assembly and moreparticularly to an improved closure assembly capable of being mounted ona container member so as to provide a portable anaerobic jar whichincludes gas generating and catalyst means thereby obviating thenecessity of providing laboratory equipment for culturing anaerobicmicroorganisms.

Anaerobe jars, such as disclosed in my prior US. Patent No. 2,463,143,granted Mar. 1, 1949, are presently in use. The anaerobe jar disclosedtherein comprises a container member having a flanged upper surface forreceiving a cast metal closure means preferably made from bronze. Theclosure means is provided with a gas inlet opening and a heating elementis suitably mounted thereon. The heating element is sealed in a brasscontainer surrounded by a plantinized catalyst wire. A fine wire safetyscreen encloses the heating element and catalyst wire. The outerperipheral edge of the cover is provided with a relatively flat, flangedsurface so that when mounted to the upper flanged surface of the jar anairtight seal is formed therebetween. To facilitate such a seal asuitable lubricant is employed.

Such an apparatus, although quite suitable for culturing anaerobicmicroorganisms in an equipped laboratory, is not completely portable,that is, it cannot be readily employed in the field where laboratoryequipment is not readily available for operating the jar assembly. Forexample, vacuum pumps, an anaerobic gas source and an electrical powersource required in operation for reacting the oxygen contained in thejar with hydrogen to provide an oxygen-free atmosphere are normally notavailable.

It is an object of the present invention to overcome the disadvantagesheretofore encountered and to provide a portable anaerobe jar assemblyhaving an improved closure therefor.

Another object of the invention is to provide an improved closureassembly which does not require inlet and outlet openings for theattachment of vacuum pumps and anaerobic gas sources. Further, it is anobject to obviate the necessity of an electrical power source forreacting with the oxygen contained in the jar with the hydrogen from theanaerobe gas source to provide an oxygen-free atmosphere.

It is another object of this invention to provide a clamping means whichwill seal the anaerobe jar yet disengage from the anaerobe jar when anexcess of gas pressure is Patented Dec. 9, 1969 present within the jarthereby preventing the anaerobe jar assembly from exploding.

A further object of this invention is to provide an assembly jar havinga catalyst for reacting with the gases in the jar and means tophysically isolate the catalyst thereby to prevent the auto-ignition ofgases present in the ar.

It is another object of this invention to provide an improved closureassembly which is readily adapted to existing anaerobe jars and which isrelatively inexpensive to manufacture and simple to operate.

My invention generally provides an improved closure asembly for mountingon an anaerobe jar for culturing anaerobic microorganisms. The anaerobejar is formed with an open upper end surrounded by an exterior flangefor the receipt of the closure assembly. The closure assembly comprisesa cover large enough to close and rest an the open upper end of theanaerobe jar and includes within the region of the cover periphery anannular cutout within which a sealing ring is disposed. The sealingring, when the cover is received on the jar, rests on the exteriorflange thereby to provide a seal between the cover and the open end ofthe jar. A clamping device for clamping the cover in sealing engagementwith the open end of the jar includes a clamping strap of a length so asto span the diameter of the cover. The clamping strap is formed withdepending end portions together with means for engaging the underside ofthe exterior flange of the jar. The clamping strap is provided with aclamping screw which is in threaded engagement with the strap and inabutting engagement with the top surface of the cover. Also, theimproved closure assembly may be provided with a relatively fine meshscreen or gauze which is mounted to the bottom surface of the cover forholding a catalyst for catalyzing the reaction between the oxygen of theatmosphere of the jar and the hydrogen or other reactants from the gassource within the jar so that the unreacted portion of the atmospherecontained in the jar is substantially oxygen free.

Other features of the invention will be best understood from thefollowing description of the drawings which illustrate the improvedanaerobe jar assembly and closure means, in which:

FIG. 1 is an elevational sectional view of my improved anaerobe jarassembly in which culture dishes are shown disposed in the jar inphantom;

FIG. 2 is a detailed elevational sectional view on an enlarged scaletaken on the lines 22 of FIG. 1 showing a housing assembly containing acatalyst therein and mounted to the under surface of the closure means;

FIG. 3 is a sectional view taken on the lines 33 of FIG. 1 showing petridishes and a disposable hydrogen generator gas source disposed in thecontainer member;

FIG. 4 is an elevational sectional view of an anaerobe jar illustratinga modified embodiment of the closure assembly and having disposedtherein a disposable anaerobic assembly; and

FIG. 5 is a partially sectional view taken on the lines 55 of FIG. 4.

Referring to the drawings and principally to FIGS. 1 and 2, the anaerobejar assembly is generally indicated by the numeral 10. Container memberor jar portion 12 is of a construction which is similar to thatconstruction of the jar as described in my US. Patent No. 2,463,143.Thus, container member 12 at its open end is provided with a flange 14having a substantially flat upper surface 15 for mounting closureassembly 16 in airtight sealing engagement thereon. The materialsemployed for making container member 12 may vary, for example, glass,rigid plastic or metal can be used. The latter two materials arepreferred when employing anaerobe jar assembly 10 for use outside of thelaboratory as a portable unit so as to reduce the possibility ofaccidentally breaking the container in transport or other handlingmishaps. However, the anaerobe jar assembly as disclosed herein iscompletely portable regardless of the materials employed for containermember 12.

My improved closure assembly 16 comprises a cover 18 having a generallyflat top surface 19. Bottom surface 20 of cover 18 is formed with adepending annular rim 21 at the periphery thereof and is provided with achannel or groove 22 for receiving a ring-shaped, replaceable, resilientwasher, gasket or O-ring 24 so that when closure assembly 16 is mountedon flat upper surface 15 of container member 12, an airtight seal isformed therebetween.

Clamping means 26 is adjustably mounted to cover 18 by threaded stud 28which is in threaded engagement with clamping means 26 through threadedopening 29. Clamping means 26 comprises: a central bridging portionhaving a pair of depending arms 30 which are bent inwardly at their ends31 so as to form a substantially U-shaped clamp. End portions 31 arepositioned under flange 14 of container member 12 so as to removablymount closure assembly 16. When threaded inwardly of the centralbridging portion, clamping means 26 will be forced in an upward positionso that ends 31 will be forced against a portion of flange 14 therebycompressing resilient member 24 so as to effect an airtight seal betweenthe cover member 18 and container member 12. Clamping means 26 ispreferably made of metal having spring-like characteristics. Further,the clamping means is sufliciently resilient so as to be yieldablethereby to allow the end portions 31 of clamping means 26 to disengagefrom flange 14 when a pressure in excess of a maximum gas pressure isformed within the anaerobe jar assembly. Thus, rupture of the containeris obviated. Therefore, the clamping means 26 is so designed as toassure absolute safety of personnel when using the assembly forculturing anaerobic microorganisms.

To help center the stud 28, a raised annular surface 32 is formed on theupper surface of cover 18 and is provided with a generally semisphericalrecess 33 to insure centering of clamping means 26.

Bottom surface 20 of cover 18 is provided with an annular raised portion34 preferably in the center of cover 18 thereby to be opposite theannular raised portion 32 on the top surface thereof. This provides asufiicient amount of material so that a threaded zone or opening 35 maybe formed therein to provide a means for mounting housing assembly 38 inwhich a catalytic agent C is disposed. Housing assembly 38, clearlyillustrated in FIG. 2, includes a cap 40 that is formed generally in theconfiguration of a truncated cone. The fiat upper surface 42 of cap 40is provided with an opening 43 to receive threaded stud 44. Threadedstud 44 may be rigidly mounted in opening 43 of cap 40 so that housingassembly 38 may be removably mounted on cover 18 in threaded opening 35.Alternatively, cover 18 may be provided with a rigidly mounted studhaving a threaded zone extending downwardly therefrom to threadedlyengage housing assembly 38 thereon. Cap 40 is provided with a generallylaterally extending annular rim 45 formed at the end portions thereof soas to provide a means for mounting a relatively fine wire mesh screen 46by simply crimping annular rim 45 in the form of a U so as to rigidlymount screen 46 therein. If desired, second screen 48 may be mounted oncap 40 at annular rim 45 along with screen 46 as shown in FIG. 2. Secondscreen 48 may or may not be of the same mesh size as screen 46 althoughsecond screen 48 is a protective screen and should generally be of asmaller mesh size so as to insure that no particles of catalyst willfall into jar 12. In addi tion, second screen 48 acts as a safety screenflame arrester. For example, if catalyst C becomes hot as a result ofcatalyzing the reaction between the oxygen present in container 12 andthe hydrogen obtained from the anaero- 4 bic gas source to form watervapor, hot spots will form on screen 46. If sufficient heat energy ispresent, the autoignition temperature of hydrogen and oxygen may bereached and cause an explosion or fire. However, screen 48 is spacedfrom screen 46 so that a temperature gradient is set up therebetweenthereby .maintaining the temperature below the auto-ignition temperatureof the gases present.

In practice, catalyst C will be disposed in screen '46 prior to mountingto cap 40. When all of the catalyst has been used, the entire housingassembly 38 is removed and catalyst C is either regenerated by standardprocedures or discarded and a new assembly inserted in its place.

Generally, it has been found that catalyst C may be provided in the formof pellets, such as 0.5% platinum on A3" alumina which is readilypurchased from Engelhard Precious Metals, Catalyst Chemical Division,Newark, New Jersey. Further, it has been found that from 5 to 50 pelletsof catalyst are sufficient to insure complete reaction of the oxygenwithin the container. Varying proportions of platinum may be applied onthe alumina if desired. Other metals may be substituted for platinum,for example, palladium, rhodium or the like. Also, the size of thepellet may vary; however, A" to A has been found to be quite suitable.

FIG. 4 illustrates a modified closure assembly 16 which is identical inconstruction to the closure assembly illustrated in FIG. 1, except thatan opening 56 is provided therein. An adapter 58 is mounted in theopening so as to provide a mounting means for collapsible expansionchamber 60. Collapsible expansion chamber 60 may be made of anyelastomeric material such as rubber or a suitable plastic which willpermit any excess gas contained in the anaerobe jar assembly to freelyexpand therein. In most instances, collapsible chamber 60 will be in theexpanded condition prior to the elimination of the oxygen contained inthe assembly.

Disposed within container 12 is an indicator assembly 50 which visuallyaids the operator of the anaerobe jar to determine when anaerobicconditions are present therein. Indicator assembly 50 comprises backingsheet 52 which is preferably made of cardboard and coated with a plasticmaterial such as polyethylene so as to render it nonabsorbent. Backingsheet 52 is preferably formed having a white background so as to readilynote a color change in indicator solution 55. Backing sheet 52 may alsobe made of a plastic material either transparent or having a whitebackground. A flexible plastic envelope 54 which is impermeable toliquid but permeable to gases is mounted to backing sheet 52. Flexibleplastic envelope 54 is preferably made of polytetrafiuoroethylene sincethis plastic material is extremely permeable to oxygen transfer. Anaqueous solution containing equal parts of 60% tri(hydroxymethyl) aminomethane, 4% dextrose and 0.02% methylene blue is placed in envelope 54and sealed. A volume of 1.5 ml. is sufiicient for most purposes.Envelope 54 may be readily mounted to backing sheet 52 by staplingthrough a tab portion formed on envelope 54 or may be mounted thereonwith a suitable adhesive. When oxygen is present in the anaerobe jar,indicator solution 55 will remain a blue-like color. However, as soon assubstantially all of the oxygen has been removed, indicator solution 55turns colorless and indicates anaerobic conditions. Methylene blue actsas a hydrogen acceptor and loses its blue color in the reduced form.When substantially all of the oxygen has been removed from the gaseousatmosphere, the indicator solution is colorless or leuco in form. Thisreaction is readily reversed by exposing indicator solution 55 to air orother oxygen sources.

After catalyst C has catalyzed the reaction between the oxygen of theatmosphere with the hydrogen from the gas source to form water vapor,the excess volume of gas will be withdrawn from collapsible expansionchamber so that while the anaerobe jar is in operation, expansion h mb r60 is in a c lapsed condition. Also, colla sible expansion chamber 60 isconstructed as a means for releasing excess pressure within the anaerobejar assembly in the event an excess amount of gas is accumulated, thusobviating the danger of the jar assembly from exploding. As statedpreviously, clamping means 26 is constructed so as to be releasable fromflange 14 of container 12 in the event an excess amount of pressure isgenerated within the anaerobe jar assembly.

When employing my improved portable anaerobe jar assembly, anaerobicmicroorganisms to be cultured are placed in container 12, usingconventional petri dishes P, as illustrated in FIG. 1 in phantom. Toprovide a ready anaerobic gas source which requires no special storagecylinders, metering valves or the like, a gas generating device G, asdisclosed in my US. Patent No. 3,246,959 and which is commerciallyavailable, may be employed.

The invention disclosed therein provides an envelope having a materialdisposed therein for evolving a predetermined amount of gas. Theenvelope is formed having a plurality of chambers. One of the chambersis provided with a material such as magnesium turnings, sodium chlorideand zinc chloride which, when mixed with a liquid, such as water,evolves or generates hydrogen as the non-toxic gas or atmospheresuitable for culturing anaerobic microorganisms. Another chamberreceives a liquid and transfers it at a controlled, relatively slow rateto the first chamber, whereby the gas or atmosphere is evolved at asimilarly controlled slow rate. In most instances, it is preferred thatthe gas evolved is hydrogen so that the hydrogen present will react withthe oxygen contained in the air in the presence of catalyst C to formwater vapor, thereby providing an anaerobic atmosphere suitable forculturing microorganisms. At the time the gas is evolving, closureassembly 16 is placed in position on container member 12. The clampingmeans 26 secures closure assembly 16 thereon in airtight relation. Sincethe amount of gas being generated is a predetermined volume, asdisclosed in my above-referenced US. patent, the possibility ofexplosion of the jar assembly is substantially eliminated.

If desirable, indicator assembly 50 may also be placed within theanaerobe jar assembly so that the operator can visually observe whenanaerobic conditions are present by the change in color of indicatorsolution 55 and also observe whether any leaks of air or other oxygensources are passing into the anaerobe jar assembly.

The anaerobe jar assembly is then placed in a thermal atmospheresuitable for the growth of anaerobic microorganisms. The thermalatmosphere is generally maintained at a temperature of 36-37 0.;however, this temperature may be varied where specific microorganismsrequire diflerent temperatures for optimum growth.

By following the above procedure for operating an anaerobe jar assemblyhaving my improved closure as sembly, it is obvious that the assembly iscompletely portable, requires no laboratory equipment for providing ananaerobic gas source which necessitates metering valves and requires nopower source for catalyzing the reaction between the oxygen present inthe container with the hydrogen or other reactants from the gas sourceto form water vapor. The anaerobe jar assembly also requires no valvesor openings in the cover assembly or container assembly for evacuatingor pressurizing the jar to provide an anaerobic atmosphere. Also, theimproved clamping means is such that it will be disengaged from theanaerobe jar so as to release the closure assembly therefrom when anexcess of pressure is present within the jar, thereby preventing anexplosion. In addition, the housing assembly for holding the catalyst isconstructed to prevent the auto-ignition of gases present in the jar dueto the heat encountered during the catalyzing of the reaction of thegases present in the jar.

The anaerobe jar with the improved cover assembly is inexpensive tomanufacture and relatively easy to construct and may be employed usingnon-technical laboratory personnel.

I claim:

1. A closure assembly for hermetically sealing a portable anaerobe jarhaving an open upper end-surrounded by an outwardly flanged annular lip,said closure assembly comprising a cover member for said jar, said covermember being of a size to rest upon and substantially completely overlaysaid outwardly flanged annular lip and including cut-out portion in theundersurface adjacent the peripheral edge adapted to receive a sealingring; a sealing ring disposed in said annular cut-out thereby to engagethe flanged annular lip surface of said anaerobe jar when said covermember is received thereon to provide an hermetic seal between the covermember and the jar; means supporting a catalytic agent on the undersideof said cover member so that when the cover is mounted on and in sealingengagement with the flanged annular lip said catalytic agent will bespatially removed from an anaerobic gas generator adapted to be disposedwithin the jar for evolving hydrogen gas yet positioned in fluid contactwith the generated gases and an oxygen atmosphere within the jar therebyto catalyze the reaction between said hydrogen gases and the oxygenatmosphere and to create anaerobic conditions within the jar whereby acharge of anaerobic microorganisms disposed within the jar may becultured, said catalytic agent supporting means including asubstantially cup shaped screen having openings of a size less than thesize of the catalytic agent therein for supporting said catalytic agentand a second cup-shaped screen in surounding spaced relation to thecatalytic agent supporting screen to provide a flame arrestor means sothat a temperature gradient is formed between the screens thereby tomaintain the temperature below the auto ignition temperature of thegases present; and clamping means for securing the cover in sealingengagement with the flanged annular jar lip, said clamping means definedby a resilient clamping strap of length to span the cover and whichincludes at each end a downward depending finger portion including meansfor engaging the undersurface of said outward flanged jar lip, saidclamping means being formed of a material having a spring-likecharacteristic so that said clamping means sealingly secures said covermember on said flanged annular lip surface yet being of sufficientresiliency so as to be yieldable and disengageable from the undersurfaceof said outward flanged lip to release the cover member from the jarupon the occurrence of internal pressures in excess of a maximumpredetermined pressure thereby to prevent damage to the jar and closuremember, and a clamping screw having an end in engagement with the topsurface of the cover and passing through the strap in threadedengagement so that said cover may be removably and sealingly mounted onsaid jar.

2. The closure assembly as set forth in claim 1 wherein said covermember is formed with an opening communicating the interior of the jarto the atmosphere and comprising a connector sealingly mounted in saidopening, and a collapsible expansion chamber supported by the connectoron the upper surface of the cover member for receiving excess atmospherecontained in said jar.

References Cited UNITED STATES PATENTS 1,137,388 4/1915 Earp-Thomas 542,279,991 4/ 1942 Hotchkiss.

2,348,448 5/ 1944 Brewer.

2,463,143 3/ 1949 Brewer 195-127 3,000,706 9/1961 Royce 195103.5 XR

ALVIN E. TANENHOLTZ, Primary Examiner US. Cl. X.R.

